中国兽医杂志2025，Vol.61Issue(4)：37-43,7.DOI:10.20157/j.cnki.zgsyzz.2025.04.005
云南省库蠓源蓝舌病病毒16型的分离鉴定
Isolation and Identification of Bluetongue Virus Serotype 16 from Culicoides(Oecacta)reconditus in Yunnan Province
摘要
Abstract
To investigate the prevalence of Culicoides-borne bluetongue virus(BTV)in certain areas of Yunnan Province,China,insect samples were collected from Yiliang County,Kunming City,and Longling County,Baoshan City between July and October 2023.Species identification and arbovirus isolation were performed.The species identification was conducted based on BLAST sequence alignment and mitochondrial cytochrome oxidase subunit I(COI)gene sequencing.The isolated arboviruses were analyzed through reverse transcription-polymerase chain reaction(RT-PCR),virus isolation and culture,indirect immunofluorescence assay(IFA),and virus titer determination,followed by genetic evolution analysis.The results showed that the collected insect samples included Culex pipiens,Culicoides reconditus,Forcipomyia bikanni,and Ceratopogonidae sp.Among them,samples collected from Longling County,Baoshan City,Yunnan Province,tested positive for BTV by RT-PCR.Following serial passage culture in BHK-21 cells,a BTV isolate was obtained.IFA confirmed that the isolate specifically reacted with BTV-16 standard positive serum,suggesting that the isolate belonged to BTV-16.This isolate was designated as YNLL-BTV16-2023,with a viral titer of 10-5.1/0.1 mL.Phylogenetic analysis revealed that the VP2 gene sequence of the isolate had the highest nucleotide homology with two Japanese isolates(GenBank accession number:LC586239.1 and AB686220.1),showing 97.20%and 96.87%similarity,respectively.The VP3 gene sequence exhibited 99.26%nucleotide homology with a Yunnan,China isolate(GenBank accession number:MG564322.1).The VP5 gene sequence had 97.77%nucleotide homology with a Japanese isolate(GenBank accession number:AB686236.1),while the VP7 gene sequence showed 98.80%nucleotide homology with two Chinese isolate(GenBank accession number:AF172831.1 and KT002584.1).These findings indicate the presence of BTV-16 strains in the border areas of Longling County,Baoshan City,Yunnan Province,with Culicoides reconditus serving as a potential vector for this serotype.This study provides valuable reference data for understanding the distribution of BTV and its vectors in the border areas of Yunnan Province.关键词
隐匿库蠓/蓝舌病病毒16型(BTV-16)/分离和鉴定/遗传进化分析Key words
Culicoides(Oecacta)reconditus/bluetongue virus serotype 16(BTV-16)/isolation and identification/phyloge-netic analysis分类
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付嘉佳,李富祥,王金萍,何于雯,谢佳芮,高华峰,李小兵..云南省库蠓源蓝舌病病毒16型的分离鉴定[J].中国兽医杂志,2025,61(4):37-43,7.基金项目
云南地方羊种质资源开发平台建设及成果转化(202102AE090039) （202102AE090039）
